Use of mitoxantrone hydrochloride liposomes for the manufacture of a medicament for treating advanced solid tumors

Abstract

Use of mitoxantrone hydrochloride liposomes in the manufacture of a medicament for treating advanced solid tumors, comprising administering to a patient with advanced solid tumor a higher therapeutically effective amount of mitoxantrone hydrochloride liposomes, e.g., 8-150 mg / m 2 The results of animal studies have shown that mitoxantrone hydrochloride liposomes can effectively inhibit the growth of various solid tumor xenografts, and the results of clinical studies have shown that mitoxantrone hydrochloride liposomes can effectively treat advanced solid tumors, and is safe and controllable.

Description

[Technical field]

[0001] The present invention claims priority to a prior application bearing patent application number 202110593815.5 and entitled "Use of Mitoxantrone Hydrochloride Liposome for Preparing Medicament for Treating Advanced Solid Tumors," filed with the State Intellectual Property Office of China on May 28, 2021, the entire text of which is incorporated herein by reference.

[0002] This patent application references PCT application WO2008 / 080367A1, filed December 29, 2007, the entire disclosure of which is incorporated herein by reference.

[0003] (Technical field) The present invention belongs to the field of antitumor medicine, specifically to the use of mitoxantrone hydrochloride liposome in the preparation of a medicament for treating advanced solid tumors. [Background technology]

[0004] Malignant tumors have become one of the major problems seriously threatening the health of Chinese people, and the incidence and mortality of malignant tumors have been increasing in the past dozen years. For most solid tumors, local tumor treatment such as surgical resection or radiotherapy is the standard treatment, but for some locally advanced or metastatic patients, when the tumor invades a wide area and is adjacent to or metastasized to important organs, it is often difficult to achieve the goal of "curing" the tumor, so chemotherapy is still the main treatment method for advanced solid tumors.

[0005] Mitoxantrone is an anthraquinone antibiotic antitumor drug, an effective inhibitor of topoisomerase II, a cell cycle non-specific drug, and has a killing effect on both hyperplastic and non-hyperplastic cancer cells in the human body. In addition, mitoxantrone can also insert into deoxyribonucleic acid through hydrogen bonds to cause cross-linking and cleavage of DNA structure, and can also bind to RNA to block the synthesis of overexpressed RNA in tumor cells, thus achieving antitumor effects. In 1987, the FDA approved NOVANTRONE (registered trademark, mitoxantrone for injection) for use in the treatment of acute myeloid leukemia, and subsequently approved multiple sclerosis, prostate cancer and other indications. According to the US drug instruction manual, for the induction treatment of acute nonlymphocytic leukemia, the combination of mitoxantrone with cytarabine is recommended, and the recommended dose of mitoxantrone is 12 mg / m2 per day. 2 is administered intravenously on days 1-3. For the treatment of multiple sclerosis, the recommended dose of mitoxantrone is 12 mg / m 2 It is administered intravenously once every 3 months. For hormone-resistant prostate cancer, the recommended dose of mitoxantrone is 12-14 mg / m 2 It is administered intravenously once every 21 days. At the same time, clinical studies have shown that injectable mitoxantrone has a certain therapeutic effect on various hematological and solid tumors, such as malignant lymphoma, breast cancer, and lung cancer, but its clinical application is very limited due to the relatively serious side effects, such as bone marrow suppression causing a decrease in white blood cells and platelets, and serious cardiac toxicity, such as palpitations, premature contractions, and electrocardiogram abnormalities.

[0006] WO2008 / 080367A1 discloses mitoxantrone liposome, and research shows that compared with the general preparation of mitoxantrone, the liposomal preparation has lower toxicity and can achieve better antitumor therapeutic effect at a relatively low dose, the entire contents of which are incorporated herein by reference.However, the above patent document does not discuss the efficacy and safety of mitoxantrone liposome in the human body.

[0007] Compared with common injections, after the drug is made into liposomes and administered to patients, its behavior in the body changes significantly, and the actual situation is very complicated. Whether the dosage is increased and the associated toxicity is permitted and acceptable is unpredictable. For example, irinotecan hydrochloride, its common injection Camptosar, when used clinically in patients with colon or rectal metastatic cancer with recurrent or progressive disease after initial fluorouracil treatment, has two dosing regimens: regimen 1 is 125 mg / m 2 1 dose per week, with a 2-week break after 4 doses. Plan 2 was 350 mg / m 2 The current clinical dosing schedule for Onivyde, a liposomal injection of irinotecan hydrochloride, is 70 mg / m 2 The dosage is 1.4 mg / m2 once every 2 weeks. As can be seen, after being formulated into liposomes, the clinical dosage is not increased, but rather significantly decreased. For example, vincristine sulfate, a commonly used injection, Oncovin, is used to treat acute leukemia, and its dosage for adults is 1.4 mg / m2. 2 is administered once a week, whereas Marqibo, a liposomal formulation of vincristine sulfate, is also administered once a week at a dose of 2.25 mg / m 2 This is 1.6 times the dosage of a typical injectable drug.

[0008] As is well known in the art, liposomes are a new drug-carrying format that can significantly change the distribution of encapsulated drugs in the body, so there are always significant differences in the safe and effective doses of liposomal formulations in the treatment of different diseases. For example, Doxil (doxorubicin hydrochloride liposome) has been approved by the FDA for three indications, which are: (1) ovarian cancer: the recommended dose is 50 mg / m 2 , intravenously once every 4 weeks; (2) Kaposi's sarcoma: The recommended dose is 20 mg / m 2 , administered intravenously once every 3 weeks; (3) Multiple myeloma: The recommended dose is 30 mg / m 2, and intravenously on day 4 after bortezomib administration. For example, AmBisome (amphotericin B liposome for injection) has the following starting doses for treating the following indications: (1) empirical treatment: recommended dose is 3 mg / kg / day, (2) system fungal infections (Aspergillus, Candida, Cryptococcus): recommended dose is 3-5 mg / kg / day, (3) cryptococcal meningitis in HIV-infected individuals: recommended dose is 6 mg / kg / day, (4) visceral leishmaniasis patients with normal immune function: 3 mg / kg / day (days 1-5), 3 mg / kg / day (days 14, 21), and visceral leishmaniasis patients with impaired immune function: 4 mg / kg / day (days 1-5), 4 mg / kg / day (days 10, 17, 24, 31, 38). As can be seen, the same drug liposome has different safe and effective doses for different indications. The dose and administration data should be individually set according to the specific disease type and the actual condition of the patient, so as to achieve the maximum efficacy and the minimum toxicity and adverse reactions, and obtain the safe and effective treatment effect of the disease.

[0009] The incidence and mortality of advanced solid tumors are both increasing, and chemotherapy is still the main treatment method at present. The therapeutic effect, safety and pharmacokinetic properties of mitoxantrone hydrochloride liposome injection in advanced solid tumors are still unclear. The present invention is to initially explore the safety and efficacy of higher doses of mitoxantrone hydrochloride liposome injection in the treatment of advanced solid tumors, so as to provide a basis for clinical application. Summary of the Invention

[0010] The present invention provides the use of mitoxantrone hydrochloride liposomes in the manufacture of a medicament for treating advanced solid tumors, preferably the use of mitoxantrone hydrochloride liposomes as the sole active ingredient in the manufacture of a medicament for treating advanced solid tumors.

[0011] In some embodiments, the drug is in an injectable form, including liquid injections, injectable powders, injectable tablets, etc. When the drug is a liquid injection, the drug contains 0.5-5 mg / mL, preferably 1-2 mg / mL, more preferably 1 mg / mL of active ingredient, calculated as mitoxantrone.

[0012] The present invention further provides a method of treating advanced solid tumors comprising administering to a patient suffering from an advanced solid tumor a therapeutically effective amount of mitoxantrone hydrochloride liposomes, preferably used alone to treat advanced solid tumors.

[0013] The present invention further provides the application of mitoxantrone hydrochloride liposomes in the treatment of advanced solid tumors.

[0014] The present invention further provides mitoxantrone hydrochloride liposomes for treating an advanced solid tumor in a patient. Preferably, the mitoxantrone hydrochloride liposomes are used alone to treat an advanced solid tumor in a patient.

[0015] In the context of the present invention, the mitoxantrone hydrochloride liposome may be in an injectable form, including liquid injection, powder for injection, tablet for injection, etc. When the drug is a liquid injection, the drug contains 0.5-5 mg / mL, preferably 1-2 mg / mL, more preferably 1 mg / mL of active ingredient, calculated as mitoxantrone.

[0016] In the context of the present invention, calculated in terms of mitoxantrone, the therapeutically effective amount is a dose of mitoxantrone hydrochloride between 8 and 150 mg / m 2 , preferably 12 to 75 mg / m 2 , and preferably 12 to 60 mg / m 2 , more preferably 16 to 54 mg / m 2 , and more preferably 20 to 49 mg / m 2 , and most preferably 24 to 40 mg / m 2The dose may be selected between any two of the above dose ranges. Specifically, when calculated using mitoxantrone, the dose is, for example, 24 mg / m 2 , 30 mg / m 2 , 36 mg / m 2 , 40 mg / m 2 , 44 mg / m 2 , 49 mg / m 2 , 54 mg / m 2 , 60 mg / m 2 , 75 mg / m 2 It is.

[0017] In the context of the present invention, the administration method of the mitoxantrone hydrochloride liposome is intravenous administration. Preferably, in the case of intravenous administration every time, the drip administration time of the liposome drug formulation is 60 minutes or more, preferably 60 to 120 minutes, more preferably 90±15 minutes. Preferably, the administration cycle is once every 3 weeks.

[0018] In the context of the present invention, the advanced solid tumor is preferably an advanced solid tumor for which conventional treatments are ineffective or for which no effective treatment is available, including situations where there is currently no standard treatment or the patient is unable to tolerate the standard treatment, such as human prostate cancer, progressive lymphoma, relapsed / refractory lymphoma, breast cancer (e.g. progressive recurrent or metastatic breast cancer), ovarian cancer, fallopian tube cancer, intestinal cancer, lung adenocarcinoma, and uterine leiomyosarcoma, preferably fallopian tube cancer, intestinal cancer, lung adenocarcinoma, and uterine leiomyosarcoma.

[0019] In the context of the present invention, the "therapeutically effective amount" refers to the amount of mitoxantrone hydrochloride liposome administered to a patient each time, which is capable of effectively treating, controlling or ameliorating advanced solid tumors in the patient.

[0020] In the context of the present invention, unless otherwise stated, the doses are calculated in mitoxantrone.

[0021] In the context of the present invention, the mitoxantrone hydrochloride liposomes can be prepared by conventional methods in the art, and may be mitoxantrone hydrochloride liposomes prepared by any one of the methods disclosed in the prior art, for example, by the method disclosed in WO2008 / 080367A1.

[0022] In some embodiments, the drug or mitoxantrone hydrochloride liposomes have one or more of the following properties: (i) the particle size of the mitoxantrone hydrochloride liposome is about 30 to 80 nm, for example, about 35 to 75 nm, about 40 to 70 nm, about 40 to 60 nm, or about 60 nm; (ii) mitoxantrone hydrochloride forms a poorly soluble precipitate with multivalent counterions (e.g., sulfate, citrate, or phosphate) in the liposome; (iii) the phospholipid bilayer in the mitoxantrone hydrochloride liposome contains a phospholipid having a phase transition temperature (Tm) higher than body temperature, so that the phase transition temperature of the liposome is higher than body temperature, for example, the phospholipid is selected from hydrogenated soybean lecithin, phosphatidylcholine, hydrogenated egg yolk lecithin, lecithin dipalmitate, lecithin distearate, or any combination thereof; (iv) the phospholipid bilayer in the mitoxantrone hydrochloride liposome contains hydrogenated soy lecithin, cholesterol, and distearoylphosphatidylethanolamine modified with polyethylene glycol 2000 (DSPE-PEG2000); (v) the phospholipid bilayer in the mitoxantrone hydrochloride liposome contains hydrogenated soybean lecithin, cholesterol, and distearoylphosphatidylethanolamine modified with polyethylene glycol 2000 in a mass ratio of about 3:1:1, mitoxantrone hydrochloride forms a poorly soluble precipitate with polyvalent acid ions in the liposome, and the particle size of the mitoxantrone hydrochloride liposome in the drug is about 60 nm; (vi) Mitoxantrone hydrochloride liposome is mitoxantrone hydrochloride liposome of the national drug regulatory standard H20220001.

[0023] In the context of the present invention, the mitoxantrone hydrochloride liposome has a particle size of about 30-80 nm and contains 1) an active ingredient mitoxantrone capable of forming a poorly soluble precipitate with a multivalent counterion in the liposome, and 2) a phospholipid bilayer containing a phospholipid having a phase transition temperature (Tm) higher than body temperature, so that the phase transition temperature of the liposome is higher than body temperature. The phospholipid having a Tm higher than body temperature is one or more selected from phosphatidylcholine, hydrogenated soybean lecithin, hydrogenated egg yolk lecithin, dipalmitic acid lecithin, and distearic acid lecithin, and the particle size is about 35-75 nm, preferably about 40-70 nm, more preferably about 40-60 nm, and particularly preferably about 60 nm. The phospholipid bilayer contains hydrogenated soybean lecithin, cholesterol, and distearoylphosphatidylethanolamine modified with polyethylene glycol 2000, in a mass ratio of 3:1:1, the particle size is about 60 nm, and the counter ion is sulfate ion. Preferably, the phospholipid bilayer of the liposome contains hydrogenated soybean lecithin, cholesterol, and distearoylphosphatidylethanolamine modified with polyethylene glycol 2000, in a mass ratio of 3:1:1, the particle size is about 40-60 nm, the counter ion is sulfate ion, and the weight ratio of HSPC:Chol:DSPE-PEG2000:Mitoxantrone in the liposome is 9.58:3.19:3.19:1.

[0024] In the context of the present invention, the method for preparing the mitoxantrone hydrochloride liposomes is as follows. HSPC (hydrogenated soybean lecithin), Chol (cholesterol), and DSPE-PEG2000 (distearoylphosphatidylethanolamine modified with polyethylene glycol 2000) were weighed in a mass ratio of 3:1:1 and dissolved in 95% ethanol to obtain a clear solution (i.e., an ethanol solution of phospholipids). The ethanol solution of phospholipids was mixed with a 300 mM ammonium sulfate solution and hydrated by shaking at 60-65°C for 1 h to obtain heterogeneous multivesicular liposomes. Then, the particle size of the liposomes was reduced using a microjet device. The obtained sample was diluted 200 times with a 0.9% NaCl solution, and then detected by NanoZS, and the average particle size of the particles was about 60 nm, with the main peak concentrated at 40-60 nm. Then, the ammonium sulfate in the blank liposome external phase was removed using an ultrafiltration device, and the external phase was replaced with 290 mM sucrose and 10 mM glycine to form a transmembrane ammonium sulfate gradient. Mitoxantrone hydrochloride solution (10 mg / mL) was added to the blank liposomes at a lipid-drug ratio of 16:1, and the drug was loaded at 60-65 °C. After about 1 h incubation, the encapsulation efficiency can be proven to be about 100% using gel exclusion chromatography. The product obtained with this formulation is named PLM 60. The weight ratio of HSPC:Chol:DSPE-PEG2000:mitoxantrone in PLM 60 is 9.58:3.19:3.19:1, and the osmotic pressure of the sucrose-glycine solution is close to the physiological value.

[0025] It should be understood that a number of technical details and parameters in the above exemplary manufacturing method can be debugged and determined within a reasonable range by those skilled in the art. For example, the types of amino acids that can replace glycine in the external phase to form a transmembrane ammonium sulfate gradient include, but are not limited to, histidine, asparagine, glutamic acid, leucine, proline, and alanine. Also, for example, the mass ratio of HSPC, Chol, and DSPE-PEG2000 can be appropriately adjusted. Furthermore, for example, for the parameters of lipid-drug ratio in the manufacture of a specific liposome drug formulation, those skilled in the art can design, measure, and finally obtain a suitable lipid-drug ratio to increase drug loading as much as possible while simultaneously reducing drug leakage. For the present mitoxantrone hydrochloride liposomal formulations, the range of lipid-drug ratios available is wide, for example, it may be as low as 2:1 or as high as 30:1, 40:1 or 50:1, with more suitable lipid-drug ratios being about (15-20):1, for example, about 15:1, 16:1, 17:1, 18:1, 19:1 or 20:1. Thus, the multiple advantageous properties of the mitoxantrone hydrochloride liposomal formulations described above are even more important, and the methods of achieving these properties are diverse. Effect of the Invention

[0026] The results of the animal test of the present invention show that mitoxantrone hydrochloride liposome injection can significantly inhibit tumor growth in a dose-dependent manner, and its therapeutic effect is much stronger than that of the general injection of mitoxantrone hydrochloride, and its duration of therapeutic effect is obviously superior to that of the latter, and it is proved that mitoxantrone hydrochloride liposome injection has a better therapeutic effect in inhibiting tumor growth and / or prolonging the survival rate of tumor-bearing animals compared with the general injection at the same dose.The pharmacokinetic test shows that mitoxantrone hydrochloride liposome has a remarkable long-term circulation property in the body, a remarkable targeting property to tumor tissue, and linear kinetic property in the body, and does not accumulate in the body during the interval between two doses, compared with the general preparation.At the same time, the results of clinical study show that mitoxantrone hydrochloride liposome can effectively treat advanced solid tumors, has a good therapeutic effect, and has few adverse reactions. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0027] The technical solution of the present invention will be described in more detail below with reference to specific examples. The following examples are merely for illustrative purposes and should not be construed as limiting the scope of the present invention. Any technology realized based on the above content of the present invention is included in the scope of the present invention.

[0028] Unless otherwise specified, all materials and reagents used in the following examples are commercially available or can be prepared by known methods.

[0029] In the present invention, the following abbreviations are used: CR: Complete response. The specific definition is based on the Response Evaluation Criteria in Solid Tumors (RECIST 1.1), which is used internationally in studies. PR: Partial response. The specific definition is based on the Response Evaluation Criteria in Solid Tumors (RECIST 1.1), which is used internationally in studies. SD: Stable disease. The specific definition is based on the Response Evaluation Criteria in Solid Tumors (RECIST 1.1), which is accepted in international studies. PD: Progression of disease. The specific definition is based on the Response Evaluation Criteria in Solid Tumors (RECIST 1.1), which is used internationally. Objective response rate (ORR) = (CR + PR) / total number of evaluable cases × 100%. Disease control rate (DCR) = (CR + PR + SD) / total number of evaluable cases × 100%. Example 1: Tumor implantation study in mice Hereinafter, Mit-Inj stands for mitoxantrone hydrochloride injection, and Mit-Lipo stands for mitoxantrone hydrochloride liposomal PLM 60.

[0030] NU / NU mice were subcutaneously inoculated with human prostate cancer PC-3 cells, and on the 28th day after inoculation, the mice were divided into groups and administered with solvent (5% glucose injection), Mit-Lipo (Shijiazhuang Zhongqi Pharmaceutical Group Co., Ltd., lot number 070501) and Mit-Inj (Chongqing Kailin Pharmaceutical Co., Ltd., lot number M20050201) by intravenous injection, twice at 6-day intervals (q6d×2), and the study was terminated on the 26th day after grouping. The experimental results showed that there was a significant difference (P<0.05) in the tumor volume over time in each treatment group. The relative tumor volume data (see Table 1, which refers to the ratio of tumor volume at a specific time point to tumor volume at the initial time point (D0)) showed that the tumor growth inhibitory effect of the Mit-Inj 2 mg / kg group was maintained until the 20th day after administration, while each Mit-Lipo group inhibited tumor growth in a dose-dependent manner, and the tumor inhibitory effect was maintained until the end of the study (day 26). Under the same dose conditions, the Mit-Lipo 2 mg / kg dose group had a significant difference (P<0.05) compared with Mit-Inj on days 20 to 26 after the first administration. During the study, one animal died in the Mit-Inj group, and none died in the other groups. This shows that compared with general injections, mitoxantrone hydrochloride liposome injection has a better tumor growth inhibitory effect, a longer maintenance time, and is safe and well tolerated.

[0031] [Table 1]

[0032] Example 2: Acute toxicity study results of mitoxantrone liposome injection The acute toxicity evaluation of single doses of mitoxantrone hydrochloride liposomes and mitoxantrone hydrochloride injection was carried out in mice and dogs. The results showed that the toxicity of mitoxantrone formulated in liposomes was significantly reduced compared to mitoxantrone hydrochloride injection.

[0033] [Table 2]

[0034] Example 3: Phase I Tolerance and Pharmacokinetics Clinical Study of Mitoxantrone Hydrochloride Liposomal Injection A randomized, open-label, single-center, dose-escalation phase I clinical study was completed between August 2011 and June 2013 to evaluate the tolerance of mitoxantrone hydrochloride liposomal injection in patients with advanced tumors and to simultaneously conduct pharmacokinetic studies. Mitoxantrone hydrochloride liposomal injection was administered at a dose of 6 mg / m 2 , 10 mg / m 2 , 12 mg / m 2 , 14 mg / m 2 , 16 mg / m 2 , 18 mg / m 2 A total of six dose groups were set up and tested, and mitoxantrone hydrochloride for injection (common formulation) 10 mg / m 2 was used as a control. Each cycle had 28 days, and the drug was administered on the first day of each cycle for a maximum of three cycles. The study included 20 patients, 17 of whom received mitoxantrone hydrochloride liposome injection and 3 of whom received injectable mitoxantrone hydrochloride (a common formulation). The study included 20 patients with the same dose level (10 mg / m 2 ) showed that the incidence of leukopenia and neutrophils in the control group (general injection of mitoxantrone hydrochloride) was significantly higher than that in the test group, and the severity was also more severe than that in the test group. No DLT reactions occurred in the mitoxantrone hydrochloride liposome test group. The evaluation results of the initial treatment effect showed that the ORR was 22.9% (2 / 9) and the DCR was 44.4% (4 / 9).

[0035] From February 2014 to September 2017, the supplementary phase I clinical trial of mitoxantrone hydrochloride liposome, "Multiple-dose tolerance clinical study of mitoxantrone hydrochloride liposome injection", was conducted at the Fourth Hospital of Hebei Medical University. The study continued to investigate the tolerance and safety of mitoxantrone hydrochloride liposome injection in patients with advanced lymphoma and explored the initial therapeutic effect. The study included 18 mg / m 2 , 20 mg / m 2 , 22 mg / m 2 , 24 mg / m2 A total of four dose groups were designed. Each cycle consisted of 28 days, and treatment was administered on the first day of each cycle for a maximum of four cycles. A total of 17 subjects were enrolled in this study. During the course of the study, 2 One DLT response was observed only in the 100 mg / kg / day dose group, and no DLTs were observed. Early efficacy results showed an ORR of 35.3% (6 / 17) and DCR of 47.4% (9 / 17).

[0036] As can be seen, the change in the formulation improved the safety and tolerability of mitoxantrone. Mitoxantrone hydrochloride liposome injection was administered at a dose of 6 to 24 mg / m 2 It is safe and tolerated within limits, and there is also room for further improvement in therapeutic efficacy with increasing doses.

[0037] Example 4: Pharmacokinetic clinical study of mitoxantrone hydrochloride liposomal injection A pharmacokinetic clinical study (HE071-07) was conducted for relapsed / refractory lymphoma subjects from July 2019. Eighteen to 24 patients were randomly enrolled in a 1:1:1 ratio, and the test doses were 12 mg / m 2 , 16 mg / m 2 , 20 mg / m 2 The drug concentrations of total mitoxantrone and free mitoxantrone in human plasma at different doses were measured for a total of 4 cycles Q4W, and the PK parameters were calculated. As of May 19, 2020, the PK characteristics of 18 patients were obtained. The results are shown in Tables 3 and 4.

[0038] 12 mg / m 2 , 16 mg / m 2 , 20 mg / m 2 The AUC of total and free mitoxantrone in plasma was 0-∞ Both increased with increasing dose, and the AUC of total mitoxantrone 0-∞ The PK results showed that the 16 mg / m 2and 20 mg / m 2 After administration of the two doses, the C max The values ​​were found to be relatively close.

[0039] [Table 3] [Table 4]

[0040] Example 5: Phase II Clinical Study of Mitoxantrone Hydrochloride Liposomal Injection in Advanced Recurrent / Metastatic Breast Cancer A randomized, controlled, single-center Phase II clinical study was conducted on patients with advanced recurrent or metastatic breast cancer, with a total of 60 breast cancer patients, including 10 patients in the experimental group (mitoxantrone hydrochloride liposome injection 20 mg / m 2 , administered once every 4 weeks) and a control group (mitoxantrone hydrochloride injection 14 mg / m 2 The study group consisted of 30 patients each (administered once every 4 weeks). The results showed that the ORR, DCR, and PFS of the study group were all superior to those of the control group. In terms of safety, the incidence of elevated troponin T, a marker of myocardial damage, in the mitoxantrone hydrochloride liposome injection group was much lower than that of the general mitoxantrone hydrochloride injection group (3.3% vs. 36.7%), demonstrating good overall cardiac safety.

[0041] Example 6: A Phase I clinical study of dose escalation and dose expansion of mitoxantrone hydrochloride liposome injection in Chinese patients with advanced solid tumors Summarizing the above research results, and referring to the consideration of the starting dose in the Technical Guidance Principles for Clinical Trials of Antitumor Drugs issued by the National Medical Products Administration, it is stated that "For cytotoxic drugs, the calculation of the starting dose for a single dose in Phase I clinical trials should, in principle, be in mg / m 2"This is equivalent to 1 / 10 of the MTD dose in rodents in non-clinical trials, or 1 / 6 of the MTD dose in non-rodents. At the same time, it is also necessary to consider the toxic reactions and reversibility of the MTD dose in different animal species. In general, it is appropriate to select the MTD of the most correlated animal to calculate the dose. When the correlation of animals is unclear, it is appropriate to select the MTD of the most sensitive animal to calculate." A phase I clinical study of dose escalation and dose expansion of mitoxantrone hydrochloride liposome injection (provided by Shijiazhuang Zhongnuo Pharmaceutical Group Co., Ltd., the formulation of which is the same as the national drug quasi-character H20220001 drug) was conducted in patients with advanced solid tumors in China. The starting dose in the study was 24 mg / m 2 The maximum dose of mitoxantrone hydrochloride liposome was determined based on the toxicity data from non-clinical studies. Pre-clinical animal toxicity studies showed that the MTD in ICR mice was 25 mg / kg, which is lower than the human MTD of 75 mg / m 2 This corresponds to an MTD of 2 mg / kg in beagle dogs and a human MTD of 40 mg / m 2 Based on this, the human MTD is 40-75 mg / m 2 Based on subject safety considerations, the 40 mg / m 2 was selected as the maximum escalating dose.

[0042] [Table 5]

[0043] First, test design This study was an open-label, multi-center, phase I clinical study, mainly divided into two stages, dose escalation and dose expansion, and included subjects with advanced solid tumors. The study was administered mitoxantrone hydrochloride liposome injection as a single agent to explore the safety and tolerance of the drug, determine the maximum tolerated dose (MTD), and observe the pharmacokinetic characteristics, while evaluating the therapeutic effect (referring to RECIST 1.1 criteria).

[0044] The study included a screening period, a treatment period, and a follow-up period. The screening period was 28 days, and subjects completed screening-related tests after signing informed consent and a baseline evaluation. Subjects who passed the screening proceeded to the treatment period. In the treatment period, subjects received monotherapy with mitoxantrone hydrochloride liposome injection, and the dosing schedule was to administer the drug once every 3 weeks on the first day of each cycle. The first cycle of the dose escalation phase was the DLT observation period, and subjects who did not experience DLT could continue to receive the next cycle according to the dose of the first cycle. For subjects who experienced DLT, the investigators could decide according to the situation. If the subjects were evaluated as continuing to tolerate the treatment and gaining benefit, they would continue to receive the drug at a lower dose level, and if the subjects were evaluated as continuing to not tolerate the treatment and not gaining benefit, they would be withdrawn from the study. In the dose expansion phase, there was no need to observe DLT, and a total of 6 cycles were administered. For subjects who completed 6 cycles of administration, if the subjects continued to gain benefit from the treatment and were tolerant, the investigators and sponsors could consider and decide whether to continue treatment. During the treatment period, the treatment effect was evaluated once every two cycles. Subjects were required to collect pharmacokinetic (PK) blood samples at different time points before and after administration according to the requirements of the plan. All subjects completed relevant examinations and safety and tolerability observations during the course of the study. An end-of-treatment visit was conducted 4 weeks after the last dose, and survival follow-up was performed once every 6 weeks thereafter (treatment effect should be evaluated for subjects who did not experience disease progression).

[0045] 1. Dose Escalation Phase The starting dose of mitoxantrone hydrochloride liposome injection was 24 mg / m 2 and a higher dose of 30 mg / m 2 , 36 mg / m 2 , 40 mg / m 2 To The study followed a "3+3" dose escalation design, where at a particular dose level, if all three patients had no dose-limiting toxicity (DLT) in the first cycle, the dose was escalated to the next dose, and if one DLT occurred in the first cycle, three more patients were included, and if three or six patients had two or more DLTs in the first cycle, dose escalation was stopped, the dose was deemed an "intolerant dose," and the dose group before that dose was defined as the maximum tolerated dose (MTD). The predefined maximum dose (40 mg / m 2 If no maximum tolerated dose is found at 4 mg / m 2, the investigator and sponsor will consider whether to continue dose escalation. Possible test doses include, for example, 44 mg / m 2. 2 , 49 mg / m 2 , 54 mg / m 2 , 60 mg / m 2 , 75 mg / m 2 , 150 mg / m 2 or a dose between any two of the dose ranges above.

[0046] For subjects who experience DLT, the investigator may make a case-by-case decision. If the subject is assessed as still benefiting from treatment, they may continue on at the next lower dose level (20 mg / m for the first dose group). 2 The dose level was then increased to 100 mg / kg / day, and if the subject was assessed to continue not benefiting from the treatment, they were withdrawn from the study. Dose levels were not allowed to be increased in the same subject.

[0047] 2. Dose-limiting toxicity (DLT) DLT was defined as any adverse event related to study drug within 21 days (D1–D21) after the first dose, as determined by NCI-CTCAE version 5.0.

[0048] The specific criteria were as follows: hematological toxicity 1) Grade 4 neutropenia not resolving within 7 days; 2) Grade 4 thrombocytopenia, decreased hemoglobin; 3) Grade 3 or greater febrile neutropenic fever that did not resolve within 3 days; 4) Grade 3 thrombocytopenia with bleeding tendency.

[0049] Other non-hematologic toxicities 1) ≥ grade 2 reduced ejection fraction; 2) Any other non-hematologic toxicity ≥ Grade 3, except in the following circumstances: <1> Transient (≤24 hours) grade 3 fever (>40°C), fatigue, headache, or nausea that resolved to grade 1 or baseline after treatment; <2> Grade 3 diarrhea, vomiting, and electrolyte disturbances (including hypokalemia, hypophosphatemia, hypocalcemia, etc.) resolved to Grade 1 or baseline within 48 hours after treatment.

[0050] 3. Dose Expansion Phase During the study, the safety and tolerability of the subjects were evaluated, and 2 to 4 groups were selected according to the subjects' conditions to carry out case expansion (for the dose group to evaluate tolerance, dose expansion can be carried out at the same time), and 10 to 20 patients were expanded in each group for the selected dose groups. Safety, tolerability, pharmacokinetic properties and efficacy were further observed. The expanded cases were not included in the DLT observation population.

[0051] II. Test procedure Each subject underwent the following study sequence: a screening period from days -28 to -1, a treatment period (6 cycles), an end-of-treatment visit, and survival follow-up.

[0052] After the subjects signed the informed consent and completed all baseline examinations during the screening period, subjects who met the enrollment criteria and did not meet the exclusion criteria were allowed to receive treatment with mitoxantrone hydrochloride liposome injection. During the treatment period, all subjects underwent tumor evaluation once every two cycles to observe the initial therapeutic effect of the study drug, and at the same time, they completed the relevant examinations specified in the plan to observe the safety and pharmacokinetic properties, and PK (pharmacokinetic) blood samples were taken at different time points before and after administration according to the plan. Subjects performed an end-of-treatment visit 28 (± 7) days after the last dose, and then followed up for survival once every 6 weeks (treatment effect needs to be evaluated for subjects who do not experience disease progression).

[0053] 3. Test group Only subjects who meet all of the following entry criteria and do not meet any of the exclusion criteria may be enrolled in this clinical study.

[0054] (1) Registration Criteria Subjects must meet all of the following criteria: 1. The subject willingly participates in the study and signs an informed consent; 2. Men and women, ages 18-65; 3.Patients with advanced solid tumors confirmed by histopathology / cytology; 4.Patients with advanced solid tumors for which conventional treatments are ineffective or lack effective treatments, including those for which there is currently no standard treatment and the patient is unable to tolerate the standard treatment, as determined by the investigator; 5. At baseline, have at least one measurable lesion meeting the RECIST 1.1 definition; 6.ECOG score of 0 to 1; 7. Resolution of prior antitumor treatment toxicity to ≤ Grade 1 (excluding alopecia, pigmentation, or other toxicity not considered to pose a safety risk to the subject in the study); 8. Adequate organ function and laboratory values ​​must meet the following requirements: Absolute neutrophil count (ANC) ≧ 1.5 × 10 9 / L (no G-CSF leukocytosis treatment within 2 weeks prior to laboratory test); ● hemoglobin (Hb) ≥ 9.0 g / dL (no red blood cell transfusion within 2 weeks prior to laboratory test); Platelets ≥ 100 x 10 9 / L (no platelet transfusion within 2 weeks prior to laboratory test); ● Creatinine ≤ 1.5 × ULN; ● Total bilirubin ≤ 1.5 × ULN; • Alanine aminotransferase (AST) / aspartate aminotransferase (ALT) ≤ 3 × ULN; ● Blood coagulation function: prothrombin time (PT), international normalized ratio (INR) ≤ 1.5 × ULN; 9. Female subjects had negative urine or blood HCG (excluding menopause and hysterectomy), and the subjects and their partners were using effective contraception during the study period and within 6 months after the last dose.

[0055] (2) Exclusion criteria Subjects who meet any of the following criteria will be excluded from this study: 1. Severe allergy to mitoxantrone or liposomal drugs; 2. Subjects with brain or meningeal metastases; 3. Survival period < 3 months; 4. Patients with chronic hepatitis B (HBsAg or HBcAb positive and HBV DNA ≥ 1000 IU / mL), chronic hepatitis C (HCV antibody positive and HCV RNA higher than the lower limit of detection at the research center), or HIV antibody positive; 5. Had an active bacterial, fungal, or viral infection requiring intravenous infusion within 1 week prior to the first drug administration; 6.Subjects who have received any anti-tumor treatment (e.g., radiotherapy, targeted therapy, immunotherapy, endocrine therapy, etc.) within 4 weeks prior to the first administration, or who have received any herbal medicine or traditional Chinese medicine approved for oncology indications within 2 weeks prior to the first administration; 7. Received treatment with other investigational drugs within 4 weeks prior to first dose; 8.Patients who have undergone major surgery within 3 months prior to the first dose and have not yet recovered from the surgery, or who are scheduled to undergo major surgery during the study period; 9. A serious thrombotic or embolic event, e.g., pulmonary embolism, occurred within 6 months prior to screening as adjudicated by the investigator; 10. Had other active malignancies within the past 3 years, except for cured locally curable cancers such as basal cell or squamous cell skin cancer or in situ prostate cancer, cervical cancer, or breast cancer; 11. Cardiac dysfunction, including any of the following: • Long QTc syndrome or QTc interval >480 ms; • Complete left bundle branch block, second- or third-degree atrioventricular block; • Severe, uncontrollable arrhythmias requiring drug treatment; • History of chronic congestive heart failure, NYHA ≥ grade 3; ● Cardiac ejection fraction (ejection fraction) less than 50% within 6 months; • valvular heart disease with CTCAE ≥ grade 3; • Uncontrolled hypertension (defined as systolic blood pressure ≥ 160 mmHg or diastolic blood pressure ≥ 100 mmHg under drug control); • Within 6 months prior to screening, there was a history of myocardial infarction, unstable angina, severe pericardial disease, or electrocardiographic evidence of acute ischemic or severe conduction system abnormalities; 12. Have received doxorubicin or other anthracycline therapy and have a cumulative doxorubicin dose of 350 mg / m 2 (anthracycline equivalent dose calculation: 1 mg doxorubicin = 2 mg epirubicin = 2 mg pirarubicin = 2 mg daunorubicin = 0.5 mg demethoxydaunorubicin = 0.45 mg mitoxantrone; doxorubicin liposomal does not calculate cumulative dose); 13. Breastfeeding women; 14. Suffered from severe and / or uncontrolled illnesses or other illnesses that, in the investigator's judgment, may affect the patient's participation in this study (including, but not limited to, diabetes mellitus that is not effectively controlled, kidney disease requiring dialysis, severe liver disease, life-threatening autoimmune and bleeding disorders, substance abuse, nervous system disorders, etc.); 15. Any other reason that makes the participant unsuitable for participation as determined by the investigator.

[0056] (4) Exit / stop criteria Subjects may withdraw from the study without reason or discontinue the study intervention at any stage of the study. The reason a subject discontinues or withdraws from the study should be recorded in the eCRF and, if possible, investigators should conduct a termination visit and evaluation for subjects who withdraw from the study.

[0057] Subjects must withdraw from the study if: • The subject withdraws informed consent or the subject or a family member requests to leave the study; ● Participated in other clinical studies during the course of the study (excluding OS follow-up).

[0058] 4. Research results This time, the therapeutic effects of 19 patients were evaluated, and according to the type of tumor, they were divided into 7 cases of breast cancer, 7 cases of ovarian cancer and fallopian tube cancer, 3 cases of intestinal cancer, 1 case of lung adenocarcinoma, and 1 case of uterine leiomyosarcoma.The results of the study show that the use of mitoxantrone hydrochloride liposome injection has the effects of sustained release, targeting, reduced toxicity and synergism after the drug enters the human body through intravenous injection, and has good prospects for clinical application in patients with multiple types of advanced solid tumors.

[0059] 24 mg / m 2 and 30 mg / m 2 No DLTs (dose-limiting toxicities) occurred in any of the dose-escalation studies. A total of 25 subjects were enrolled, with 2 15 patients in the 30 mg / m 2Adverse reactions were primarily hematological toxicity, with 10 patients in the 24 mg / m 2 Grade 3 AEs occurring in >5% of patients in the 30 mg / m 2 Grade 3 AEs with an incidence of >5% in the 24 mg / m group were neutropenia (30%), leukopenia (30%), thrombocytopenia (20%), anemia (10%), and lymphocyte count decrease (10%). 2 In the group, 12 patients had at least one efficacy evaluation, 2 had PR, 4 had SD, and 6 had PD, giving an ORR of 16.7% and DCR of 50%. 2 Seven patients in the group had at least one therapeutic effect evaluation, four had SD, and three had PD, resulting in a DCR of 57.1%. Patients who were evaluated as SD continued to receive the drug, leading to better therapeutic effects.

[0060] The incidence of all-grade adverse reactions of mitoxantrone hydrochloride liposome injection in this study compared with the generic formulation of mitoxantrone included leukopenia (76% vs 87%), neutropenia (64% vs 79%), thrombocytopenia (32% vs 39%), anemia (60% vs 75%), and lymphocyte count decline (4% vs 72%), and cardiac safety included cardiac dysfunction (0% vs 18.0%), myocardial ischemia (0% vs 5.0%), and arrhythmia (0% vs 7.0%). Mitoxantrone safety data were extracted from the CALGB 9182 study, which included a total of 112 subjects with advanced hormone-refractory prostate cancer and included mitoxantrone monotherapy (12-14 mg / m2) in combination with hydrocortisone. 2 ) for treatment.

[0061] As can be seen, mitoxantrone hydrochloride liposome did not show increased hematological toxicity compared with the general formulation of mitoxantrone under the condition of increasing dose, and the incidence of cardiac-related adverse reactions of the improved mitoxantrone hydrochloride liposome was obviously reduced.

[0062] The above is an illustrative description of the embodiments of the technical solution of the present invention. It should be understood that the scope of the present invention is not limited to the above embodiments. Any modifications, equivalent replacements, improvements, etc. made by those skilled in the art within the scope of the spirit and principle of the present invention should be included in the scope of the claims of this application.

Claims

1. Use of mitoxantrone hydrochloride liposomes in the manufacture of a medicament for treating an advanced solid tumor selected from ovarian or fallopian tube cancer in a patient, said mitoxantrone hydrochloride liposomes having a particle size of 30-80 nm and containing 1) an active ingredient mitoxantrone capable of forming a poorly soluble precipitate with a multivalent counter ion within the liposome, wherein said counter ion is sulfate ion, and 2) a phospholipid bilayer comprising hydrogenated soy lecithin, cholesterol, and distearoylphosphatidylethanolamine modified with polyethylene glycol 2000 in a mass ratio of 3:1:1; administering to said patient a therapeutically effective amount of said mitoxantrone hydrochloride liposomes, said therapeutically effective amount being 24-40 mg / m2.

2. The use according to claim 1, wherein the mitoxantrone hydrochloride liposome is the only active ingredient in the drug.

3. The use according to claim 1, wherein the therapeutically effective amount of the mitoxantrone hydrochloride liposome is 30 mg / m2.

4. The use according to claim 1, wherein the drug is in an injectable form, including liquid injection, powder for injection, and tablet for injection.

5. The use according to claim 4, wherein the medicament contains 0.5 to 5 mg / mL of the active ingredient, calculated as mitoxantrone.

6. The use described in claim 1, wherein the particle size is approximately 40 to 60 nm and the weight ratio of HSPC:Chol:DSPE-PEG2000:Mitoxantrone in the liposome is 9.58:3.19:3.19:

1.

7. 2) administering to said patient a therapeutically effective amount of said mitoxantrone hydrochloride liposome drug formulation, said therapeutically effective amount being 24-40 mg / m2.

8. The mitoxantrone hydrochloride liposomal drug formulation of claim 7, wherein the therapeutically effective amount of the mitoxantrone hydrochloride liposome is 30 mg / m 2 .

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